Table 1.
Two Gauss distributions of deconvoluted stacks of GP images
| Cell type (condition) | Center 1 | Width 1 | Center 2 | Width 2 | Coverage 2, % |
|---|---|---|---|---|---|
| RAW (living) | 0.18 (0.011) | 0.29 (0.055) | 0.52 (0.0059) | 0.17 (0.038) | 44.2 (6.9) |
| hMDM (living) | 0.26 (0.011) | 0.21 (0.095) | 0.58 (0.076) | 0.14 (0.042) | 29.1 (6.1) |
| THP-1 (living) | 0.11 (0.0086) | 0.41 (0.028) | 0.57 (0.028) | 0.23 (0.046) | 35.2 (10.4) |
| RAW (fixed) | 0.17 (0.063) | 0.22 (0.022) | 0.42 (0.014) | 0.25 (0.042) | 32.6 (4.4) |
| RAW (mCD-Chol) | 0.18 (0.031) | 0.23 (0.031) | 0.48 (0.010) | 0.12 (0.064) | 58.9 (7.5) |
| RAW (mCD) | 0.11 (0.011) | 0.32 (0.038) | 0.39 (0.013) | 0.15 (0.027) | 37.3 (6.1) |
Living RAW264.7, hMDM, and phorbol 12-myristate-differentiated THP-1 cells were imaged at 22°C (1.2 × 106 cells/3.14 cm2). Living RAW264.7 cells were pretreated at 22°C with 1.0 mg/ml mCD for 10 min or 0.5 mg/ml cholesterol-complexed mCD (mCD-Chol) for 1 h. Where indicated, Laurdan-labeled (30 min, 0.5 μM) RAW264.7 cells were fixed in 4% paraformaldehyde at 4°C for 20 min. Stacks of GP images, as shown in Fig. 1, were fitted to two Gauss distributions, and the centers, widths, and percentage of pixels associated with center 2 (“coverage”) were averaged over 8-15 stacks.